Inhibitory properties of Enterococcus spp. isolated from faeces of healthy dogs

The aim of the present study was to evaluate the inhibitory effect by the cross-streak method of nine Enterococcus faecium strains isolated from faeces of healthy dogs and their treated and non-treated cell-free supernatant (CFS) by the well-diffusion test on the growth of potentially pathogenic bacteria isolated from clinical cases and aflatoxigenic Aspergillus section Flavi and the consequent aflatoxin B₁ (AFB₁) production. Results obtained from the cross-strake assay showed that E. faecium MF1, GJ18 and GJ40 presented the major inhibitory activity against all pathogenic strains assayed; E. faecium GJ40 produced the larger inhibitory zones (26–27 mm). Well-diffusion test results showed that the majority of the enterococci strains CFS had antimicrobial activity against the pathogenic microorganisms, especially on Gram negative indicators. Cell-free supernatant of E. faecium GJ40 was the one that produced the largest inhibition zones (14 to 21 mm) in the majority of the indicator microorganisms assayed. All supernatants treated with 10 N NaOH (pH6) showed no inhibitory effect on the indicator strain assayed. With respect to fungal inhibition, any of the CFS assayed significantly inhibited the Aspergillus strains growth. But, in general, all CFS reduced AFB₁ production from 8 to 87%. The results demonstrate that enterococci isolated from healthy dog feaces produce substances with the capacity to inhibit some potential pathogenic bacteria growth and the capacity of inhibiting or reducing the AFB₁ production in vitro.     

Influence of herbicide glyphosate on growth and aflatoxin B1 production by Aspergillus section Flavi strains isolated from soil on in vitro assay

The effect of six glyphosate concentrations on growth rate and aflatoxin B₁ (AFB₁) production by Aspergillus section Flavi strains under different water activity (a_W) on maize-based medium was investigated. In general, the lag phase decreased as glyphosate concentration increased and all the strains showed the same behavior at the different conditions tested. The glyphosate increased significantly the growth of all Aspergillus section Flavi strains in different percentages with respect to control depending on pesticide concentration. At 5.0 and 10 mM this fact was more evident; however significant differences between both concentrations were not observed in most strains. Aflatoxin B₁ production did not show noticeable differences among different pesticide concentrations assayed at all a_W in both strains. This study has shown that these Aspergillus flavus and A. parasiticus strains are able to grow effectively and produce aflatoxins in high nutrient status media over a range of glyphosate concentrations under different water activity conditions.     

14C‐labeled aflatoxin B1 prepared with yeastlike cultures of Aspergillus parasiticus

Abstract

A simple method was developed to produce ¹⁴C‐labeled aflatoxin B₁ by using the yeastlike phase of Aspergillus parasiticus NRRL 2999. Yeastlike cultures resulted from absence of manganese in a synthetic medium. Sodium acetate‐1‐¹⁴C had a 0.22% average incorporation; sodium acetate‐1,2‐¹⁴C, 0.70%. The average yield of labeled B₁ was 10 mg/500 ml medium with an average specific activity of either 63.3 mCi/mole (C‐l label) or 194.3 mCi/mole (C‐1, 2 label).     

Isolation of culturable mycobiota from agricultural soils and determination of tolerance to glyphosate of nontoxigenic Aspergillus section Flavi strains

Glyphosate-based herbicides are extensively used in Argentina's agricultural system to control undesirable weeds. This study was conducted to evaluate the culturable mycobiota [colony forming units (CFU) g^?1 and frequency of fungal genera or species] from an agricultural field exposed to pesticides. In addition, we evaluated the tolerance of A. oryzae and nontoxigenic A. flavus strains to high concentrations (100 to 500 mM – 17,000 to 84,500 ppm) of a glyphosate commercial formulation. The analysis of the mycobiota showed that the frequency of the main fungal genera varied according to the analyzed sampling period. Aspergillus spp. or Aspergillus section Flavi strains were isolated from 20 to 100% of the soil samples. Sterilia spp. were also observed throughout the sampling (50 to 100%). Aspergillus section Flavi tolerance assays showed that all of the tested strains were able to develop at the highest glyphosate concentration tested regardless of the water availability conditions. In general, significant reductions in growth rates were observed with increasing concentrations of the herbicide. However, a complete inhibition of fungal growth was not observed with the concentrations assayed. This study contributes to the knowledge of culturable mycobiota from agricultural soils exposed to pesticides and provides evidence on the effective growth ability of A. oryzae and nontoxigenic A. flavus strains exposed to high glyphosate concentrations in vitro.     

Aflatoxin decomposition in various soils

Abstract

The persistence of aflatoxin in the soil environment could potentially result in a number of adverse environmental consequences. To determine the persistence of aflatoxin in soil, ¹⁴C‐labeled aflatoxin B₁, was added to silt loam, sandy loam, and silty clay loam soils and the subsequent release of ¹⁴CO₂ was determined. After 120 days of incubation, 8.1% of the original aflatoxin added to the silt loam soil was released as CO₂ ? Aflatoxin decomposition in the sandy loam soil proceeded more quickly than the other two soils for the first 20 days of incubation. After this time, the decomposition rate declined and by the end of the study, 4.9% of the aflatoxin was released as CO_2. Aflatoxin decomposition proceeded most slowly in the silty clay loam soil. Only 1.4% of aflatoxin added to the soil was released as CO₂ after 120 days incubation. To determine whether aflatoxin was bound to the silty clay loam soil, aflatoxin B₁ was added to this soil and incubated for 20 days. The soil was periodically extracted and the aflatoxin species present were determined using thin layer chromatographic (TLC) procedures. After one day of incubation, the degradation products, aflatoxins B₂ and G₂, were observed. It was also found that much of the aflatoxin extracted from the soil was not mobile with the TLC solvent system used. This indicated that a conjugate may have formed and thus may be responsible for the lack of aflatoxin decomposition.     

The in vitro effect of selected essential oils on the growth and mycotoxin production of Aspergillus species

The aim of the present study was to assess the antifungal and anti-toxinogenic activity of 15 essential oils (EOs) against three fungi of the genus Aspergillus (A. parasiticus KMi-227-LR, A. parasiticus KMi-220-LR and A. flavus KMi-202-LR). The minimum inhibitory doses (MIDs) of the tested essential oils and their antifungal activity were determined using the micro-atmosphere method. The original commercial essential oil samples of Jasminum officinale L., Thymus vulgaris L., Syzygium aromaticum (L.) Merrill &; Perry, Rosmarinus officinalis L., Ocimum basilicum L., Eucalyptus globulus Labill., Salvia officinalis L., Citrus limon (L.) Burm, Origanum vulgare L., Lavandula angustifolia Mill., Carum carvi L., Citrus sinensis (L.) Osbeck., Zingiber officinalis Rosc., Mentha piperita L. and Cinnamomum zeylanicum Nees. (C. verum J.S.Presl.) were produced in Slovakia (Calendula a.s., Nová ?ubovňa, Slovakia). All essential oils exhibited activity against all tested strains of fungi. After 14 days of incubation, A. flavus (KMi-202-LR) showed the highest susceptibility with a growth inhibition percentage (GIP) of 18.70% to C. limon and 5.92% to C. sinensis, while A. parasiticus (KMi-220-LR) exhibited a GIP of 20.56% to J. officinale. The minimum inhibitory doses (MIDs) of EOs with the most significant activity were recorded. The best antifungal activity, using the micro-atmosphere method was found in S. aromaticum with an MID of 62.5 μL L^?1 air, T. vulgaris (MID of 62.5 μL L^?1 air) and O. vulgare (MID of 31.5 μL L^?1 air) against all tested strains. Mycotoxin production of the tested strains was evaluated by the thin layer chromatography (TLC) method. Mycotoxin production of AFB₁ and AFG₁ was inhibited following all treatments with C. carvi, R. officinale and S. officinale, Eucalyptus globulus L. and O. basilicum L. Essential oils exhibited a potential inhibition activity against toxic fungi, although, these affected only the production of AFB₁.     

Evaluation of Saccharomyces cerevisiae strains as probiotic agent with aflatoxin B1 adsorption ability for use in poultry feedstuffs

In this study the aflatoxin B₁ (AFB₁) removal capacity, the tolerance to salivary and gastrointestinal conditions, autoaggregation and coaggregation with pathogenic bacteria of Saccharomyces cerevisiae strains isolated from broiler feces, were evaluated. Only four of twelve isolated strains were identified as Saccharomyces cerevisiae using molecular techniques. The results obtained in AFB₁ binding studies indicated that the amount of AFB₁ removed was both strain and mycotoxin-concentration dependent. Therefore, a theoretical model was applied in order to select the most efficient strain to remove AFB₁ in a wide range of mycotoxin concentration. The results indicated that S. cerevisiae 08 and S. cerevisiae 01 strains were the most efficient microorganisms in the mycotoxin removal. Viability on simulated salivary and gastrointestinal conditions was investigated and S. cerevisiae 08 strain showed the best results, achieving 98% of total survival whereas S. cerevisiae 01 reached only 75%. Autoaggregation and coaggregation assays showed S. cerevisiae 08 as the most appropriate strain, mainly because it was the unique strain able to coaggregate with the four bacterial pathogens assayed. Consequently, S. cerevisiae 08 is the best candidate for future in vivo studies useful to prevent aflatoxicosis. Further quantitative in vitro and in vivo studies are required to evaluate the real impact of yeast-binding activity on the bioavailability of AFB₁ in poultry. However, this study could be useful in selecting efficient strains in terms of AFB₁ binding and provide an important contribution to research into microorganisms with potential probiotic effects on the host.     

Antimicrobial potential of commercial silver nanoparticles and the characterization of their physical properties toward probiotic bacteria isolated from fermented milk products

The application of nanotechnology in the agriculture and food sector is relatively recent compared to its usage in drug delivery or pharmaceuticals. Therefore, this paper presents a study of the effect of silver nanoparticles on probiotic bacteria based on the example of Lactobacillus acidophilus LA-5, Bifidobacterium animalis subsp. lactis BB-12 and Streptococcus thermophilus ST-Y31 isolated from fermented milk products. Probiotic bacteria are one of the most crucial groups of bacteria for the food industry, because of their claimed health-promoting properties. Studies have shown that the type and concentration of silver nanoparticle solutions have a significant impact on the tested probiotic bacteria which are profitable for the digestive system. In the presence of all tested silver nanoparticles, St. thermophilus ST-Y31 growth was inhibited significantly by the dilution method as opposed to the disk-diffusion method. Both the disk-diffusion and the dilution methods showed no significant differences between L. acidophilus LA-5 and B. animalis subsp. lactis BB-12. The concentrations 2 μg mL^?1 and 0.25 μg mL^?1 had the highest antibacterial activity and statistically significant impacts on the tested probiotic strains. To our knowledge, this is the first report on potential antimicrobial effect of nanosilver against the health-promoting probiotic bacteria L. acidophilus LA-5, B. animalis subsp. lactis BB-12 and St. thermophilus ST-Y31 isolated from fermented milk products.     

Fullerol C<Subscript>60</Subscript>(OH)<Subscript>24</Subscript> nanoparticles and mycotoxigenic fungi: a preliminary investigation into modulation of mycotoxin production

Increased use of fullerols in various fields and expected increase of their environmental spread impose the necessity for testing fullerol nanoparticles (FNP) effects on microbiota. There is little information available on the interaction of mycotoxigenic fungi and FNP, despite FNP having a great potential of modifying mycotoxin production. Namely, FNP exhibit both ROS-quenching and ROS-producing properties, while oxidative stress stimulates mycotoxin synthesis in the fungi. In order to shed some light on the extent of interaction between FNP and mycotoxigenic fungi, the effects of fullerol C₆₀(OH)₂₄ nanoparticles (10, 100, 1000 ng/mL) on mycelial growth, aflatoxin production and oxidative stress modulation in an aflatoxigenic strain of Aspergillus flavus (NRRL 3251) during 168 h of incubation in a liquid culture medium were examined. FNP slightly reduced mycelial biomass weight, but significantly decreased aflatoxin concentration in media. Lipid peroxide content, superoxide dismutase, catalase and glutathione peroxidase activities suggest that FNP treatments hormetically reduced oxidative stress within fungal cells in turn suppressing aflatoxin production. These findings contribute to the assessment of environmental risk and application potential of fullerols.     

Fungal levels in houses in the Fukushima Daiichi Nuclear Power Plant evacuation zone after the Great East Japan Earthquake

Residences located within 20 km of the damaged Fukushima Daiichi Nuclear Power Plant were evacuated shortly after the Great East Japan Earthquake. The levels of airborne and surface fungi were measured in six houses in the evacuation zone in August 2012 and February 2013. Airborne fungal levels in all of the houses in the summer were higher than the environmental standard levels for residential houses published in Architectural Institute of Japan (>1000 colony-forming units [CFU]/m³). In two houses whose residents rarely returned to visit, fungal levels were extremely high (>52,000 CFU/m³). Although fungal levels in the winter were much lower than those in the summer, they were still higher than environmental standard levels in several houses. Indoor fungal levels were significantly inversely related to the frequency with which residents returned, but they were not correlated with the air exchange rates, temperature, humidity, or radiation levels. Cladosporium spp. and Penicillium spp. were detected in every house. Aspergillus section Circumdati (Aspergillus ochraceus group) was also detected in several houses. These fungi produced ochratoxin A and ochratoxin B, which have nephrotoxic and carcinogenic potential. The present study suggests that further monitoring of fungal levels is necessary in houses in the Fukushima Daiichi Nuclear Power Plant evacuation zone, and that some houses may require fungal disinfection.

Implications: The results suggest that residents’ health could be at risk owing to the high levels of airborne fungi and toxic fungi Aspergillus section Circumdati. Therefore, monitoring and decontamination/disinfection of fungi are strongly recommended before residents are allowed to return permanently to their homes. In addition, returning to home with a certain frequency and adequate ventilation are necessary during similar situations, e.g., when residents cannot stay in their homes for a long period, because fungal levels in houses in the Fukushima Daiichi Nuclear Power Plant evacuation zone were inversely correlated with the frequency with which residents returned to visit their houses.     

Characterization of chlordecone-tolerant fungal populations isolated from long-term polluted tropical volcanic soil in the French West Indies

The insecticide chlordecone is a contaminant found in most of the banana plantations in the French West Indies. This study aims to search for fungal populations able to grow on it. An Andosol heavily contaminated with chlordecone, perfused for 1 year in a soil–charcoal system, was used to conduct enrichment cultures. A total of 103 fungal strains able to grow on chlordecone-mineral salt medium were isolated, purified, and deposited in the MIAE collection (Microorganismes d'Intérêt Agro-Environnemental, UMR Agroécologie, Institut National de la Recherche Agronomique, Dijon, France). Internal transcribed spacer sequencing revealed that all isolated strains belonged to the Ascomycota phylum and gathered in 11 genera: Metacordyceps, Cordyceps, Pochonia, Acremonium, Fusarium, Paecilomyces, Ophiocordyceps, Purpureocillium, Bionectria, Penicillium, and Aspergillus. Among predominant species, only one isolate, Fusarium oxysporum MIAE01197, was able to grow in a liquid culture medium that contained chlordecone as sole carbon source. Chlordecone increased F. oxysporum MIAE01197 growth rate, attesting for its tolerance to this organochlorine. Moreover, F. oxysporum MIAE01197 exhibited a higher EC₅₀ value than the reference strain F. oxysporum MIAE00047. This further suggests its adaptation to chlordecone tolerance up to 29.2 mg l^?1. Gas chromatography–mass spectrometry (GC-MS) analysis revealed that 40 % of chlordecone was dissipated in F. oxysporum MIAE01197 suspension culture. No chlordecone metabolite was detected by GC-MS. However, weak amount of ¹⁴CO₂ evolved from ¹⁴C₁₀-chlordecone and ¹⁴C₁₀-metabolites were observed. Sorption of ¹⁴C₁₀-chlordecone onto fungal biomass followed a linear relationship (r ²?=?0.99) suggesting that it may also account for chlordecone dissipation in F. oxysporum MIAE01197 culture.     

Degradation of Metolachlor in Crude Extract of Aspergillus Flavus

Abstract

Crude enzyme from a soil fungus, Aspergillus flavus, was isolated from a field soil following repeated applications of metolachlor [2-Chloro-N-(methoxy-1-methylethyl)-2′-ethyl-6′-methyl acetanilide]. Metolachlor hydrolysis by the crude enzyme extract was determined by enzyme assay. The tests were performed in phosphate buffer, pH 7.5, and the reaction was carried out at two herbicide concentrations (20 and 100 μg mL^?1) and two crude extract volumes (0.2 and 0.5 mL of the homogenized crude extract mixture). The rate of metolachlor degradation was found faster in samples containing higher volume of crude extract, (T _1/2, 5.7 h) for both concentrations of the herbicide. The activities of enzymes responsible for dechlorination coupled with hydroxylation, N-dealkylation, and breaking of amide linkage were found responsible in the degradation.     

Potentiation of antifungal effect of a mixture of two antifungal fractions obtained from Baccharis glutinosa and Jacquinia macrocarpa plants

The aim of the present work was to evaluate the effect of mixtures of antifungal fractions extracted from Baccharis glutinosa and Jacquinia macrocarpa plants on the development of the filamentous fungi Aspergillus flavus and Fusarium verticillioides. The minimal inhibitory concentration that inhibited 50% of growth (MIC₅₀) of each plant antifungal fraction was determined from the percentage radial growth inhibition of both fungi. Binomial mixtures made with both plant fractions were used at their MIC₅₀ to determine the Fractional Inhibitory Concentration index (FIC index) for each fungus in order to evaluate their synergistic effect. Each synergistic mixture was analyzed in their effect on spore germination, spore size, spore viability, mitotic divisions, hyphal diameter and length, and number of septa per hypha. Some antifungal mixtures, even at low concentrations, showed higher antifungal effect than those of the individual antifungal fraction. The FIC indices of mixtures that showed the highest antifungal activity against A. flavus and F. verticillioides were 0.5272 and 0.4577, respectively, indicating a synergistic effect against both fungi. Only 12% and 8% of the spores of A. flavus and F. verticillioides, respectively, treated with the synergistic mixtures, were able to germinate, although their viability was not affected. An increase in the number of septa per hypha of both fungi was observed. The results indicated that the synergistic mixtures strongly affected the fungal growth even at lower concentrations than those of the individual plant fractions.     

Mechanism of aflatoxin uptake in roots of intact groundnut (Arachis hypogaea L.) seedlings

Aflatoxins are one of the most potent toxic substances that occur naturally, which enter agricultural soils through the growth of aflatoxigenic fungi in rhizhosphere and nonrhizhosphere soils. Though several reports regarding the uptake of aflatoxin by plants are available, the mechanism of aflatoxin uptake remains unknown. This study characterized the aflatoxin uptake mechanism by in vitro hydroponic experiments under variable conditions. The uptake reached saturation after 48 h of incubation for AFB₁ and B₂ and 60 h for AFG₁ and G₂. A linear increase in uptake with increasing aflatoxin concentrations was observed, and it fits both linear and nonlinear regression. AFB₁ uptake was directly proportional to transpiration rate, and blocking aquaporin activity using mercuric chloride revealed its involvement in the uptake. None of the metabolic inhibitors used to block active transport had any effect on aflatoxin uptake except for sodium azide. From the present study, it could be concluded that aflatoxin uptake by groundnut roots followed mainly a passive way and is facilitated through aquaporins. The involvement of active component should be studied in detail.     

Rate of iodine volatilization and accumulation by filamentous fungi through laboratory cultures

Five strains of basidiomycetes (Lentinula edodes, Coprinus phlyctidosporus, Hebeloma vinosophyllum, Pleurotus ostreatus and Agaricus bisporus), one strain of ascomycete (Hormoconis resinae) and six strains of imperfect fungi (Penicillium chrysogenum, Penicillium roquefortii, Cladosporium cladosporioides, Alternaria alternata, Aspergillus niger and Aspergillus oryzae) were cultured in a liquid medium containing a radioactive iodine tracer (¹²⁵I), and were tested for their abilities to volatilize or accumulate iodine. Of the fungal strains tested, 11 strains volatilized a considerable amount of iodine, with L. edodes showing the highest volatilization rate of 3.4%. The volatile organic iodine species emitted from imperfect fungi cultures was identified as methyl iodide (CH₃I). In contrast, six fungal strains in 12 strains accumulated a considerable amount of iodine from the medium with concentration factors of more than 1.0. Among these, Alt. alternata and Cl. cladosporioides accumulated more than 40% of the iodine in their hyphae, and showed high concentration factors of 22 and 18, respectively. These results suggest that filamentous fungi have a potential to influence the mobility and speciation of iodine by volatilization and accumulation. Considering their great biomass in soils, filamentous fungi may contribute to the global circulation of stable iodine and also the long-lived radioiodine, ¹²⁹I (half-life: 1.6 × 10⁷ years), released from nuclear facilities into the environment.     

Assessment of indoor air in Austrian apartments with and without visible mold growth

Fungal spores are transported across great distances in the outdoor air and are also regularly found indoors. Building conditions and behavior-related problems in apartments may lead to massive growth of mold within a very short period of time.The aim of this study was to evaluate whether the visible growth of mold indoors influences the concentration of fungal spores in the air as well as the variety of their species. Samples were collected from 66 households in Austria. For each sampling, the corresponding outdoor air was measured as reference value. The size of the visible mold growth was categorized in order to correlate the extent of mold growth with the concentration of airborne spores as well as the fungal genera. In order to determine fungal spore concentrations in the air, the one-stage MAS-100^® air sampler was used. Malt extract agar (MEA) and dichloran glycerol agar (DG18) plates were used as culture media. The total colony forming units (CFU) per m³ were determined. The fungi were identified from the isolated colonies.The results show that in apartments visibly affected by mold, the median values were significantly higher than those of apartments without visible mold growth. The extent of visible mold growth is significantly correlated with both concentration of fungal spores (p<0.001) as well as the predominance of Penicillium sp. and Aspergillus sp. (p<0.001) in indoor air. The total fungal concentration of Penicillium and Aspergillus in the air of apartments is recommended for assessing fungal exposure.     

In vitro effects of various xenobiotics on Fusarium spp. strains isolated from cereals

This study aimed to determine the susceptibility of Fusarium spp. strains isolated from cereals to selected heavy metals, fungicides and silver nanoparticles. The experiments were conducted using 50 Fusarium strains belonging to five species: F. graminearum, F. culmorum, F. oxysporum, F. sporotrichioides and F. avenaceum. The strains were found to be highly resistant to Pb²⁺ and Zn²⁺. Medium resistance to Cu²⁺ and Mn²⁺ and low resistance to Cd²⁺ and Fe³⁺ was also observed. Among the tested fungicides, formulations containing azoxystrobin, prochloraz and tebuconazole proved to be the most effective in inhibiting the growth of fungi, as they affected fungal growth in each of the applied doses. Susceptibility of Fusarium spp. to nanosilver, demonstrated in this study, shows the legitimacy of using nanostructures as fungicidal agents. The results confirm high diversity of the analyzed fungal species in terms of susceptibility to the tested substances, and encourage to continue research on the resistance of Fusarium spp. to various fungicidal agents.     

Factors affecting the removal of aflatoxin M1 from food model by Lactobacillus and Bifidobacterium strains

This paper describes the ability of six dairy strains of Lactobacillus and Bifidobacterium to remove aflatoxin M₁ (AFM₁) from phosphate-buffered saline (PBS) and reconstituted milk. Bacteria were incubated in both PBS and reconstituted milk containing 5, 10 and 20 ng mL^?1 for 0, 4 and 24 h at 37°C. After centrifugation the concentration of AFM₁ was determined in the supernatant fraction using high-performance liquid chromatography. The binding abilities of AFM₁ by viable (10⁸ CFU mL^?1) and heat-killed Lactobacillus and Bifidobacterium strains in PBS ranged from 10.22 to 26.65% and 14.04 to 28.97%, respectively. Similarly, AFM₁-binding capacity in reconstituted milk was found to range from 7.85 to 25.94% and from 12.85 to 27.31% for viable and heat-killed bacteria, respectively within 4 h. While B. bifidum Bb 13 was the best binder, the poorest removal was achieved by L. acidophilus NCC 68. Binding was reversible, and a small proportion of AFM₁ was released back into the solution. The toxin concentration and incubation period had no effect on the removal of AFM₁ by bacteria both in PBS and reconstituted milk.     

Occurrence of aflatoxin M1 in human milk samples in Vojvodina,Serbia: Estimation of average daily intake by babies

The objectives of the study were to determine the aflatoxin M1 content in human milk samples in Vojvodina, Serbia, and to assess the risk of infants' exposure to aflatoxins food contamination. The growth of Aspergillus flavus and production of aflatoxin B1 in corn samples resulted in higher concentrations of AFM1 in milk and dairy products in 2013, indicating higher concentrations of AFM1 in human milk samples in 2013 and 2014 in Serbia. A total number of 60 samples of human milk (colostrum and breast milk collected 4–8 months after delivery) were analyzed for the presence of AFM1 using the Enzyme Linked Immunosorbent Assay method. The estimated daily intake of AFM1 through breastfeeding was calculated for the colostrum samples using an average intake of 60 mL/kg body weight (b.w.)/day on the third day of lactation. All breast milk collected 4–8 months after delivery and 36.4% of colostrum samples were contaminated with AFM1. The greatest percentage of contaminated colostrum (85%) and all samples of breast milk collected 4–8 months after delivery had AFM1 concentration above maximum allowable concentration according to the Regulation on health safety of dietetic products. The mean daily intake of AFM1 in colostrum was 2.65 ng/kg bw/day. Results of our study indicate the high risk of infants' exposure, who are at the early stage of development and vulnerable to toxic contaminants.